id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-304421-xpj6c0vx	Piñón, Josefina D.	Further Requirements for Cleavage by the Murine Coronavirus 3C-like Proteinase: Identification of a Cleavage Site within ORF1b	1999-10-25		.txt	text/plain	7343	346	57	The coronavirus 3C-like proteinase (3CLpro), flanked on either side by hydrophobic, possibly membrane-spanning regions (HD1 and HD2), is believed to be the prinicipal viral proteinase responsible for the processing events leading to the formation of the viral replicase complex, with as many as 11 potential cleavage sites identified throughout pp1ab (Gorbalenya et al., 1989; Lee et al., 1991 ) (see Fig. 1 ). We therefore created several substrates of various lengths, encoding different putative cleavage sites in ORF1b, in order to investigate processing by the recombinant MBP-3CLpro. The effect of these cleavage site mutations on the autocatalytic cis release of the 29-kDa 3CLpro was assayed by the expression of the Radiolabeled, in vitro transcribed, and translated substrate from pET21-NX.3C was incubated with MBP-3CLpro (lane 2) or an equal volume of column buffer/20% glycerol (lane 1) and the processed products were separated on a 15% SDS-PAGE gel.	./cache/cord-304421-xpj6c0vx.txt	./txt/cord-304421-xpj6c0vx.txt